Studies On The Preparation Of BCB And Its Pharmacodynamics

Boswellia carterii Birdwood (Frankincense) is used for promoting blood flow, inducing menstruation to stop pain, detumescence and promoting granulation in traditional Chinese medicine. Modern pharmacological study shows that boswellic acids can resist proliferation, induce differentiation and apoptosis of tumor cells and that the extract of the oleogum resin of B. carterii can induce the apoptosis of ANLL cells, APL HL60 cells, AMCL U937 cell and AML primary cell. B.carterii extractive is also an inhibitor of topoisomerase I and II. However, the evaporable components can seriously stimulate the gastrointestinal tract. So, the compliance of the patients is poor.The research was aimed to systematically study the preparation of B.carterii volatile oil liposomes and solidify the preparation parameters; To establish the quality evaluation system and the methodologies of measuring the drug content and entrapment efficiency; To establish a noumenal animal model to compare the anti-tumor effect of B.carterii volatile oil, BCB liposomes and a positive control drug—5-Fu.The first part describes our research on the measurement and methodologies of the major quality evaluation standard of liposomes—entrapment efficiency. On the condition that we do not know the composition of B.carterii volatile oil, for the first time, UV and GC spectrophotometries was adopted to measure the drug content. In order to estimate the entrapment of BCB in liposomal formulations, we developed extraction method which is seldomly applied to separate the unentrapped drug from liposomes, which is very important to the future studies on formulation.According to the characteristics of volatile oil, we adopted a liposome preparation method which is called alcohol-dripping and transonic to prepare BCB containing liposomes in this paper and whole process can be easily applied into industrial manufacturing. The preparation parameters and formulation were optimized based on the above-mentioned quality evaluation method. In order to ensure high entrapment efficiency, the orthogonal design was carried out and the optimum formulation of liposome was obtained. The ideal conditions were found out to be: Lecithin : cholesterol: 6 : 1, Lecithin : drug : 6 : 1, preparation temperature : 60℃. The average entrapment efficiency of the optimized BCB liposomes is 87.7±1.23%.The quality of BCB liposomes was evaluated by the appearance, the particle size, pH, Zeta potential, stability, entrapment efficiency and percolation rate. And the shape of BCB liposomes was observed by transmission electron microscopy. The results demonstrated this preparation has reached the original requirements of design.Later, human hepatocelluar cancer(HepG2) was established in nude mouse model. This helped to investigate the anti-hepatocelluar cancer effect of BCB liposomes in vivo. The results indicated that BCB liposomes and 5-Fu injection significantly inhibited the growth of hepatocelluar cancer in nude mice. And BCB liposomes at a middle dose (187.5μg·10g^-1, i.p) could effectively inhibit the growth of tumor and have the similar efficiency with 5-Fu injection which has already been applied in clinic. And also, the pharmacological efficiency to the tumor depended on the drug concentration. The animal tolerance experiment included hematology and marrow test. The experiment showed that the tolerance of the animal was good. Utilizing the pathological technology, we compared HE staining of drug and the negative,positive control groups. Meanwhile, the expression of PCNA, VEGF, CD-31 in HepG2 was studied based on immunohistochemical technology. The research demonstrated that BCB and its liposomes have strong inhibitive effect to HepG2.The research successfully established the methodologies of measuring the drug content and entrapment efficiency and quality evaluation system. Applying the orthogonal design, we screened the best formulation and preparation parameters. Balb/c-nu animal model showed BCB liposomes and 5-Fu injection significantly inhibited the growth of hepatocelluar cancer in nude mice. The pharmacological efficiency to the tumor depended on the drug concentration. Immunohistochemical results demonstrated that BCB and its liposomes have strong inhibitive effect to HepG2.